RAM-induced allostery facilitates assembly of a Notch pathway active transcription complex
RAM-induced allostery facilitates assembly of a Notch pathway active transcription complex
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DOI：
10.1074/jbc.m709501200
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发表时间：
2008-05-23
影响因子：
4.8
通讯作者：
Kovall, Rhett A.
中科院分区：
文献类型：
--
作者：
Friedmann, David R.;Wilson, Jeffrey J.;Kovall, Rhett A.
Notch通路是一种保守的细胞间信号传导机制，其中细胞外信号通过核效应器CSL被转导成转录输出。CSL通过形成CSL-NotchIC-Mastermind三元复合物从阻遏物转化为激活物。NotchIC的RAM（RBP-J相关分子）结构域与CSL强烈相互作用;然而，其在CSL-NotchIC-Mastermind三元复合物组装中的作用尚不清楚。在这里，我们提供了一个全面的热力学，结构和生化分析的RAM-CSL相互作用的组件从小鼠和蠕虫。我们的结合数据表明，RAM和CSL形成一个高亲和力的复合物在DNA的存在或不存在。我们的结构研究揭示了一个显着的远端构象变化CSL RAM结合后，这创造了一个对接网站的Mastermind结合到复杂的。最后，我们表明，在反式RAM肽的加入有利于在体外形成的CSL-NotchIC-Mastermind三元复合物。
The Notch pathway is a conserved cell-to-cell signaling mechanism, in which extracellular signals are transduced into transcriptional outputs through the nuclear effector CSL. CSL is converted from a repressor to an activator through the formation of the CSL-NotchIC-Mastermind ternary complex. The RAM (RBP-J associated molecule) domain of NotchIC avidly interacts with CSL; however, its role in assembly of the CSL-NotchIC-Mastermind ternary complex is not understood. Here we provide a comprehensive thermodynamic, structural, and biochemical analysis of the RAM-CSL interaction for components from both mouse and worm. Our binding data show that RAM and CSL form a high affinity complex in the presence or absence of DNA. Our structural studies reveal a striking distal conformational change in CSL upon RAM binding, which creates a docking site for Mastermind to bind to the complex. Finally, we show that the addition of a RAM peptide in trans facilitates formation of the CSL-NotchIC-Mastermind ternary complex in vitro.